Table 3.

Phenotypic plate assays

Phenotypic test	Reagent(s) used	Concn tested	Phenotypic observations
Role of pesL in siderophore biosynthesis	Iron stresses (high, low, and none)	10 μM, 1.5 mM, and 200 μM BPSb	NDa
Oxidative stress	Menadione	20, 30, and 40 μM	The A. fumigatus ΔpesL mutant is more resistant to menadione at all of the concentrations tested
	Diamide	0.1, 0.2, 0.4, 1, and 2 mM	ND
	Hydrogen peroxide	1, 2, and 3 mM	The ΔpesL and Δpes1 mutants display increased sensitivity to H2O2
Antifungal susceptibility	Voriconazole	0.05-0.25, 0.5, 0.75, and 1.0 μg/ml	The ΔpesL and Δpes1 mutants display increased sensitivity to voriconazole
	Amphotericin B	0.125, 0.25, 0.5, and 1.0 μg/ml	The ΔpesL and Δpes1 mutants display increased sensitivity to amphotericin B
	Caspofungin	0.2, 0.5, and 1.0 μg/ml	ND
Heavy metal stress	Cobalt chloride	0.1, 0.5, and 1 mM	ND
Cell wall stress	Caffeine	2 and 5 mM	ND
	Congo Red	5, 10, and 15 μg/ml	ND
	Calcofluor White	100 and 200 μg/ml	ND
	High temp (48°C)	NA	ND
Membrane stress	SDS	0.01 and 0.02% (wt/vol)	ND

^aND, no difference between wild-type and NRPS mutants.

^bBPS, bathophenanthroline disulfonate.